嗜热脂肪芽孢杆菌质粒DNA的高压电穿孔转化研究  

STUDY ON GENETIC TRANSFORMATION OF BACILLUS STEAROTHERMOPHILUS WITH PLASMID DNA BY ELECTROPORATION

作　　者：陈兰明[1] 盛祖嘉[1] 毛裕民[1] 

机构地区：[1]复旦大学遗传学研究所,上海200433

出　　处：《微生物学通报》2000年第2期85-89,共5页Microbiology China

基　　金：国家自然科学基金!No.39230020

摘　　要：采用高压电穿孔法将穿梭质粒导入了嗜热脂肪芽胞杆菌（Ｂａｃｉｌｌｕｓ　ｓｔｅａｒｏｔｈｅｒｍｏｐｈｉｌｕｓ）Ｋ１０４１和Ｔ５２１菌株。以对数生长后期的菌体制备Ｋ１０４１转化细胞，以ＬＢ平板上于５０℃培养的过夜菌制备Ｔ５２１转化细胞，细胞密度为５～７×１０９细胞／ｍＬ。电击条件如下：电容２５μＦ，电场强度１０．０ＫＶ／ｃｍ，脉冲控制器设定２００。 Ｋ１０４１和 Ｔ５２１最高转化效率分别达２．０１×１０和１．１９ ×１０转化子／μｇ ＤＮＡ。此外，研究发现Ｔ５２１和Ｋ１０４１中存在着ＤＮＡ的限制／修饰系统。Transformation of shuttle plasmids into Bacillus stearothermophilus K1041 and T521 by electroporation was carried out. K1041 cells for transformation were prepared from late-logarithmic growth phage, T521 cells prepared after growth on LB plate at 50℃ overnight. The cell concentration was about 5-7 × 10cells/mL. The electroporation conditions were to set capacitor at 25μF, electrical field strength at 10.0KV / cm and the pulse controller at 200 ohms. The maximum transformation efficiencies of K1041 and T521 were up to 2.01 × 104 and 1.19 × 102 transformants/μg DNA, respechvely. In addition,evidence indicated that T521 and K1041 possessed DNA restriction and modification systems.

关 键 词：嗜热脂肪芽胞杆菌 质粒 电击转化 

分 类 号：Q933[生物学—微生物学]